Primary and secondary metabolism, and post-translational protein modifications, as portrayed by proteomic analysis of Streptomyces coelicolor
Primary and secondary metabolism, and post-translational protein modifications, as portrayed by proteomic analysis of Streptomyces coelicolor
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DOI：
10.1046/j.1365-2958.2002.03219.x
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发表时间：
2002-11-01
影响因子：
3.6
通讯作者：
Chater, KF
中科院分区：
文献类型：
--
作者：
Hesketh, AR;Chandra, G;Chater, KF
新测序的天蓝色链霉菌基因组估计编码7825个理论蛋白质。我们已经用双向凝胶电泳和基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOF)实验绘制了大约10%的理论蛋白质组。鉴定了来自770个不同基因的产物，并根据其注释的功能类别讨论了所代表的蛋白质类型。平均每个基因有1.2个蛋白质，表明广泛的翻译后调控。用质谱仪对N-乙酰化、腺苷酸化和蛋白水解改性的实例进行了表征。来自初级代谢途径和某些次级代谢途径的蛋白质在图谱上都有很强的表达，并且在不止一个二维凝胶位置上鉴定了一些这些酶。因此，翻译后修饰机制可能在这些途径的调控中发挥重要作用。出乎意料的是，合成放线菌素聚酮抗生素的一种酶似乎位于细胞质隔间之外的细胞壁基质内。在编码次生代谢酶的20个基因簇中，有8个出现在蛋白质组图上，其中3个指定了新代谢物的产生。这一信息将对新代谢物的表征很有价值。
The newly sequenced genome of Streptomyces coelicolor is estimated to encode 7825 theoretical proteins. We have mapped approximately 10% of the theoretical proteome experimentally using two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. Products from 770 different genes were identified, and the types of proteins represented are discussed in terms of their annotated functional classes. An average of 1.2 proteins per gene was observed, indicating extensive post-translational regulation. Examples of modification by N-acetylation, adenylylation and proteolytic processing were characterized using mass spectrometry. Proteins from both primary and certain secondary metabolic pathways are strongly represented on the map, and a number of these enzymes were identified at more than one two-dimensional gel location. Posttranslational modification mechanisms may therefore play a significant role in the regulation of these pathways. Unexpectedly, one of the enzymes for synthesis of the actinorhodin polyketide antibiotic appears to be located outside the cytoplasmic compartment, within the cell wall matrix. Of 20 gene clusters encoding enzymes characteristic of secondary metabolism, eight are represented on the proteome map, including three that specify the production of novel metabolites. This information will be valuable in the characterization of the new metabolites.